Use of the polymerase chain reaction to detect Mycobacterium leprae in urine

Caleffi, Katiany Rizzieri; Hirata, Rosário Dominguez Crespo; Hirata, Mário Hiroyuki; Caleffi, Edilainy Rizzieri; Siqueira, Vera Ld; Cardoso, Rosilene Fressatti

doi:10.1590/s0100-879x2012007500011

Cited by 29 publications

(19 citation statements)

References 16 publications

(29 reference statements)

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“…It is clear that effective control of leprosy requires, together with multidrug therapy, new diagnostic tools that can detect M. leprae infection at an early stage. The PCR technique has been evaluated for DNA detection of M. leprae in different systems [ 18 – 20 ].…”

Section: Discussionmentioning

confidence: 99%

High frequency of M. leprae DNA detection in asymptomatic household contacts

et al. 2018

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BackgroundCharacterization of the Mycobacterium leprae genome has made possible the development of Polymerase Chain Reaction (PCR) systems that can amplify different genomic regions. Increased reliability and technical efficiency of quantitative PCR (qPCR) makes it a promising tool for early diagnosis of leprosy. Index cases that are multibacillary spread the bacillus silently, even before they are clinically diagnosed. Early detection and treatment could prevent transmission in endemic areas.MethodsIn this study, the qPCR technique is used to detect DNA of M. leprae in samples of slit skin smears (SSS) of the ear lobe and blood of leprosy patients and their asymptomatic household contacts residing in Governador Valadares, MG, Brazil, a hyperendemic area for leprosy. A total of 164 subjects participated in the study: 43 index cases, 113 household contacts, and, as negative controls, 8 individuals who reported no contact with patients nor history of leprosy in the family. The qPCR was performed to amplify 16S rRNA fragments and was specifically designed for M. leprae.ResultsOf asymptomatic household contacts, 23.89% showed bacillary DNA by qPCR in samples of SSS and blood. Also, 48.84% of patients diagnosed with leprosy were positive for qPCR while the bacillary load was positive in only 30.23% of patients. It is important to note that most patients were already receiving treatment when the collection of biological material for qPCR was performed. The level of bacillary DNA from household contacts was similar to the DNA levels detected in the group of paucibacillary patients.ConclusionConsidering that household contacts comprise a recognizable group of individuals with a high risk of disease, as they live in close proximity to a source of infection, qPCR can be used to estimate the risk of progress towards leprosy among household contacts and as a routine screening method for a chemoprophylactic protocol.Electronic supplementary materialThe online version of this article (10.1186/s12879-018-3056-2) contains supplementary material, which is available to authorized users.

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Section: Discussionmentioning

confidence: 99%

High frequency of M. leprae DNA detection in asymptomatic household contacts

et al. 2018

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“…Another method targets a gene that encodes an antigenic 36 kDa protein known as proline-rich antigen (pra gen) by cPCR [28][29][30][31][32][33], and can detect up to a single bacteria in the sample [28]. The pra gen has also been amplified by multiplex PCR (mPCR) [34]. Plikaytis and coworkers, developed a nPCR that amplifies a heat shock protein 65 kDa called groEL, which can detect 3 fg of M. leprae-DNA (single bacteria) [35].…”

Section: Leprae Detection In Clinical Samplesmentioning

confidence: 99%

“…The use of urine as a diagnosis sample is currently being evaluated [34,96]. The possibility of amplifying the M. leprae DNA in a simple non-invasive sample can be useful and maybe improve the diagnosis mainly in the tuberculoid form or in inconclusive cases where the slit skin smear is usually negative (PNL, IL and PB patients) [34].…”

Section: Urinementioning

confidence: 99%

Leprosy Diagnosis: An Update on the Use of Molecular Tools Lucrecia

Soto¹,

Muñoz²

2015

Mol Biol

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Leprosy is a chronic infectious disease caused by an obligatory intracellular mycobacteria Mycobacterium leprae, which presents tropism for Schwann cells and skin macrophages. Leprosy is a public health problem and early diagnosis is essential to avoid incapacities. The disease´s clinical presentation varies from few to widespread lesions and its diagnosis continues to be a challenge due to the low sensibility of the conventional methods, based on bacillary counts of skin smears and histopathology. Molecular techniques, especially the methods to identify M. leprae DNA based on polymerase chain reaction (PCR) have emerged as a support of the conventional methods for the analysis of clinical samples in difficult to diagnose cases, such as pure neural leprosy, indeterminate and paucibacillary leprosy. The technique has also proved useful in the study of leprosy transmission and monitoring résistance to the WHO recommended Multidrug treatment. Different biological samples can be analysed and there is no consensus in the molecular diagnostic techniques respect of the most efficient nucleic acid extraction method, most appropriate methodology and genetic target for PCR. These methods provide a very valuable option for confirmation of difficult clinical cases with scarce bacilli but requires a well-equipped laboratory and the high cost makes it inaccessible to be used as a routine diagnostic tool in most endemic countries.

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“…Polymerase chain reaction (PCR) has been used to detect M. leprae in lymph (Phetsuksiri, et al, 2006), blood (Almeida, 2004), nasal swab (Almeida, 2004), tissue fragment (Phetsuksiri, et al, 2006;Martinez, 2006), and urine (Caleffi et al, 2012(Caleffi et al, , 2010Parkash et al, 2004) clinical samples. Different PCR primers have been used to amplify the DNA of M. leprae from clinical samples (Kang, Kim, Lee, 2003).…”

Section: Introductionmentioning

confidence: 99%

Critical analysis: use of polymerase chain reaction to diagnose leprosy

Maltempe

Baldin

Lopes

et al. 2016

Braz. J. Pharm. Sci.

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Leprosy is a neglected tropical disease and an important public health problem, especially in developing countries. It is a chronic infectious disease that is caused by Mycobacterium leprae, which has a predilection for the skin and peripheral nerves. Although it has low sensitivity, slit-skin smear (SSS) remains the conventional auxiliary laboratory technique for the clinical diagnosis of leprosy. Polymerase chain reaction (PCR) is a molecular biology technique that holds promise as a simple and sensitive diagnostic tool. In the present study, the performance of two PCR methods, using different targets, PCR-LP and PCR-P, were compared with SSS with regard to leprosy diagnosis in a reference laboratory. M. leprae DNA was extracted from 106 lymph samples of 40 patients who had clinical suspicion of leprosy. The samples were subjected to both PCR techniques and SSS. Amplification of the human b-globin gene was used as PCR inhibitor control. The specificity of both PCR techniques was 100%, and sensitivity was 0.007 and 0.015 µg/ml for PCR-LP and PCR-P, respectively. No significant difference was found between either the PCR-LP or PCR-P results and SSS results (p > 0.05). Although PCR is not yet a replacement for SSS in the diagnosis of leprosy, this technique may be used as an efficient auxiliary tool for early detection of the disease, especially in endemic regions. This strategy may also be useful in cases in which SSS results are negative (e.g., in paucibacillary patients) and cases in which skin biopsy cannot be performed. A hanseníase é uma doença tropical negligenciada e ainda um importante problema de saúde pública, especialmente nos países em desenvolvimento. É uma doença infecciosa crônica causada pelo Mycobacterium leprae, que tem predileção pela pele e nervos periféricos. Embora com baixa sensibilidade, o esfregaço de linfa (SSS) continua sendo o método laboratorial convencional auxiliar no diagnóstico clínico da hanseníase. A biologia molecular representada pela Reação em Cadeia da Polimerase (PCR) trouxe a expectativa de ser uma ferramenta diagnóstica simples e sensível. No presente estudo, o desempenho de dois métodos de PCR usando alvos diferentes, PCR-P e PCR-LP, foi comparado com SSS no diagnóstico da hanseníase em um laboratório de referência. DNA de M. leprae foi extraído de 106 amostras de linfa de 40 pacientes que apresentavam suspeita clínica de hanseníase. As amostras foram submetidas tanto a PCR como SSS. A amplificação do gene humano β-globina foi usada como controle de inibição da PCR. A especificidade de ambas as técnicas de PCR foi de 100% e a sensibilidade foi de 0,007 μg/mL e 0,015 μg/mL para a PCR-P e PCR-LP, respectivamente. Não se observou diferença estatística entre os resultados da PCR-LP e PCR-P, quando comparado com SSS (p > 0,05). Apesar de a PCR ainda não substituir o SSS no diagnóstico da hanseníase, esta técnica pode ser usada como ferramenta auxiliar eficiente para a detecção precoce da doença, especialmente em regiões endêmicas. Esta estratégia pode também ser útil nos cas...

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Use of the polymerase chain reaction to detect Mycobacterium leprae in urine

Cited by 29 publications

References 16 publications

High frequency of M. leprae DNA detection in asymptomatic household contacts

High frequency of M. leprae DNA detection in asymptomatic household contacts

Leprosy Diagnosis: An Update on the Use of Molecular Tools Lucrecia

Critical analysis: use of polymerase chain reaction to diagnose leprosy

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